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Epigenetic role of epidermal growth factor expression and signalling in embryonic mouse lung morphogenesis☆

机译:表皮生长因子的表达和信号传导在胚胎小鼠肺形态发生中的表观遗传学作用☆

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Amajorunsolvedproblemindevelopmentalbiologyistodeterminewhenandhowtime-andposition-restrictedinstructionsaresignaledandreceivedduringsecondaryembryonicinductionssuchasbranchingmorphogenesis.Themouseembryoniclungrudimentwasusedtotestthehypothesisthatendogenouspeptidegrowthfactors,specificallyepidermalgrowthfactor(EGF),serveasinstructiveepigeneticsignalsformorphogenesis.ThepresenceofEGFprecursormRNAtranscriptswasdetectedusingthereverse-transcriptase-coupledpolymerasechainreactionbothinE11-E17-daymouseembryolungtissuesinvivoandinE11-daylungculturedforupto7daysinvitrounderchemicallydefined,serum-freeconditions.Immunolocalizationidentifiedaposition-restricteddistributionofEGFinandaroundtheprimitiveairwaysbothduringinvivolungmorphogenesisandinculture.EGFreceptors(EGFR)coimmunolocalizedwithEGFintheprimitiveairways.AdditionofexogenousEGFtolungsincultureresultedinsignificantconcentration-dependentstimulationofbranchingmorphogenesis,DNA,RNA,andproteincontent,andin[3H]thymidineincorporationintoDNA.Conversely,theadditionoftyrphostin(specificEGFreceptorkinaseantagonist)tolungsincultureresultedinconcentration-dependentinhibitionofbranchingmorphogenesis,DNA,RNA,andproteincontent,andin[3H]thymidineincorporationintoDNAwithoutapparentcytotoxicity.TheinhibitionoftheEGFsignalbytyrphostinwasconfirmedbyimmunoprecipitationoftyrosinephosphoproteins.WeconcludethatearlymouseembryolungsexpressEGFtranscriptsandcorrespondingEGFpeptidesinaspecificposition-restricteddistributionwhichcoimmunolocalizeswithEGFRintheprimitiveairways,whilestimulatoryandinhibitorystudiesindicateafunctionalroleforthetransducedEGFsignalintheepigeneticregulationoflungbranchingmorphogenesis.WespeculatethatthepeptidegrowthfactorEGFservesafunctioninsecondaryembryonicmorphogeneticinductions,whichmaybemodulatedbyinteractionwithothergrowthfactors.
机译:Amajorunsolvedproblemindevelopmentalbiologyistodeterminewhenandhowtime-及位置restrictedinstructionsaresignaledandreceivedduringsecondaryembryonicinductionssuchasbranchingmorphogenesis.Themouseembryoniclungrudimentwasusedtotestthehypothesisthatendogenouspeptidegrowthfactors,specificallyepidermalgrowthfactor(EGF),serveasinstructiveepigeneticsignalsformorphogenesis.ThepresenceofEGFprecursormRNAtranscriptswasdetectedusingthereverse酶 - coupledpolymerasechainreactionbothinE11-E17-daymouseembryolungtissuesinvivoandinE11-daylungculturedforupto7daysinvitrounderchemicallydefined,血清freeconditions.Immunolocalizationidentifiedaposition-restricteddistributionofEGFinandaroundtheprimitiveairwaysbothduringinvivolungmorphogenesisandinculture.EGFreceptors(EGFR)coimmunolocalizedwithEGFintheprimitiveairways.AdditionofexogenousEGFtolungsincultureresultedinsignificantconcentration-dependentstimulationofbranchingmorphogenesis,DNA,RNA,andproteincontent和[3H]胸苷掺入DN A.Conversely,theadditionoftyrphostin(specificEGFreceptorkinaseantagonist)tolungsincultureresultedinconcentration-dependentinhibitionofbranchingmorphogenesis,DNA,RNA,andproteincontent,andin [3H] thymidineincorporationintoDNAwithoutapparentcytotoxicity.TheinhibitionoftheEGFsignalbytyrphostinwasconfirmedbyimmunoprecipitationoftyrosinephosphoproteins.WeconcludethatearlymouseembryolungsexpressEGFtranscriptsandcorrespondingEGFpeptidesinaspecificposition-restricteddistributionwhichcoimmunolocalizeswithEGFRintheprimitiveairways,whilestimulatoryandinhibitorystudiesindicateafunctionalroleforthetransducedEGFsignalintheepigeneticregulationoflungbranchingmorphogenesis.WespeculatethatthepeptidegrowthfactorEGFservesafunctioninsecondaryembryonicmorphogeneticinductions,whichmaybemodulatedbyinteractionwithothergrowthfactors。

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